Construction and application of innovation gene-edited rats and intestinal 3D organoids models in drug metabolism and pharmacokinetics / 中国临床药理学与治疗学

Drug metabolism and pharmacokinetics (DMPK) are the science to study the process of drug absorption, distribution, metabolism and excretion. It is very important to evaluate the characteristics of DMPK for the early development of drugs and the later clinical precision medication. The innovative construction of DMPK models promotes the development and improvement of drug evaluation system. Based on our research results, this review summarized the latest progress and application of innovative DMPK models, focusing on the following two aspects: (1) CRISPR/Cas9 gene editing rat models, including Cyp2e1

The research progress of microRNA on regulation of drug disposition in vivo / 中国药理学通报

The disposition of drug in vivo is subjected to a series of biotransformation and transport, depending on the involvement of drug-metabolizing enzymes and transporters.However, the individual capacity varies when metabolizing and transporting the same drug, and pharmacogenomics has trouble in completely explaining the differences.microRNA, a key aspect of epigenetic modifications, is a powerful complement to traditional genetics.Emerging evidences have confirmed that drug-metabolizing enzymes and transporters be controlled by different microRNAs, and the same microRNA also regulates several drug-metabolizing enzymes or/and transporters simultaneously.All of these researches infer that microRNAs are likely to realize the comprehensive macro-regulation of gene expression.The further study of microRNAs maybe a suitable point to research the interindividual variability in disposition of drugs, and it provides a theoretical basis for rational use of drug and individualized medicine.

Screening study for genetic polymorphisms affecting pharmacokinetics of talniflumate

Talniflumate is a phthalidyl ester of niflumic acid, which has potent analgesic and anti-inflammatory effects and is widely used to treat inflammatory disorders, such as rheumatoid arthritis. To screen the possible genetic factors affecting the pharmacokinetics (PK) of talniflumate, 23 male Korean volunteers were enrolled from two separate bioequivalence studies. All subjects received 740 mg (two tablets) talniflumate in a standard 2×2 cross-over model in a randomized order. For the genetic study, PK parameters of the reference drug were used. We used Illumina Human610Quad v1.0 DNA Analysis BeadChip for whole genome single nucleotide polymorphism (SNP) analysis and whole genome genotyping data were processed by linear regression analysis for PK parameters. Whole genome analysis revealed 1498 significant SNPs (P < 0.0001) for Cmax, 65 significant SNPs (P < 0.0001) for T(max), and 1491 significant SNPs (P < 0.0001) for AUC(inf). For clinical pharmacological purposes, we selected SNPs from drug metabolizing enzymes and transporters, and analyzed the PK parameters of various genotypes. Two SNPs (rs11165069 from ABCA4 (p=0.00002); rs17847036 from CYP2C9 (p=0.000001)) showed significant associations with talniflumate C(max). In the T(max) group, two SNPs (rs3787555 from CYP24A1 (p=0.00035); rs2275034 from ABCA4 (p=0.000587)) showed significant associations with talniflumate T(max). In the AUC(inf) group, two SNPs (rs11165069 from ABCA4 (p=0.00002); rs12461006 from SLC1A6 (p=0.00008)) exhibited significant associations with talniflumate absorption. These results show that genetic factors could affect the PK parameters, and provide information that may be used in the development of personalized talniflumate therapy.

Effect of Zuotai and HgS on gene expression of drug-metabolizing enzymes in livers of mice / 中国中药杂志

Zuotai and cinnabar(96%HgS) are contained in many traditional medicines. To examine their potential effects on drug metabolism genes, mice were orally given Zuotai or HgS at doses of 10, 30, 100, 300 mg•kg⁻¹ for 7 days. HgCl2(33.6 mg•kg⁻¹) was gavaged for control. Twenty-four hour later after the last administration, livers were collected, and expressions of genes related to metabolic enzymes and transporters were examined. Zuotai and HgS had no effects on major phase-1, phase-2 and transporter genes; HgCl2 increased the expressions of CYP2B10, CYP4A10, OATP1A4, UGT1A1, UGT2A3, SULT1A1, SULT2A1, MRP1, MRP3 and MRP4; expression of OATP1A1 was decreased by HgCl2, but not by Zuotai and HgS. Therefore, Zuotai and HgS have different adverse effects on drug-metabolizing genes from HgCl2.

Screening study for genetic polymorphisms affecting pharmacokinetics of pioglitazone

Pioglitazone is known to have antidiabetic effects through decreasing peripheral, hepatic and vascular insulin resistance by the stimulation of PPAR gamma. To address the possible genetic factors affecting the pharmacokinetics (PK) of pioglitazone, 27 male Korean volunteers were enrolled from two separate bioequivalence studies. Each subject was administered 15 mg pioglitazone and reference drug PK parameters were used. We used Illumina Human610 Quad v1.0 DNA Analysis BeadChip for whole genome SNPs analysis and whole genome genotyping data was processed by linear regression analysis for PK parameters. We found 35 significant SNPs (P < 0.0001) in C(max), 1,118 significant SNPs (P < 0.0001) in T(max) and 1,259 significant SNPs (P < 0.0001) in AUC(inf) from whole genome analysis. For clinical pharmacological purpose, we selected SNPs from several phase I and II drug metabolizing enzyme and analyzed PK parameters with genotypes. Four SNPs (rs7761731 and rs3799872 from CYP39A1; rs156697 from GSTO2; rs1558139 from CYP4F2) showed significant associations with pioglitazone C(max). In the T(max) group, seven SNPs from 3 genes (rs3766198 from CYP4B1; rs2270422 from GSTZ1; rs2054675, rs10500282, rs3745274, rs8192719, and rs11673270 from CYP2B6) had significant associations. In the AUC(inf) group, seven SNPs from 4 genes (rs11572204 from CYP2J2; rs4148280 from UGT2A1, rs4646422 from CYP1A1; rs3745274, rs8192719, rs11673270, and rs707265 from CYP2B6) showed significant associations with pioglitazone absorption. These results showed that genetic makeup could affect the PK parameters and these informations could be provide information for personalized pioglitazone therapy.

Study on the polymorphisms of CYP3A5 and GSTP1 genes in the prediction of short-term efficacy of docetaxel plus thiotepa for patients with metastatic breast cancer / 中国药学杂志

OBJECTIVE: To investigate whether the polymorphisms of cytochrome P450 3A5 (CYP3A5) gene and glutathione S-transferase P1 (GSTP1) gene are associated with the short-term efficacy of docetaxel plus thiotepa for Chinese patients with metastatic breast cancer. METHODS: All patients received docetaxel plus thiotepa chemotherapy. CYP3A5 and GSTP1 were genotyped by matrix assisted laser desorption ionization / time of flight (MALDI-TOF). Disease control rate (DCR) was evaluated every two chemotherapy cycles, and was compared between different genotypes. RESULTS: Among 93 enrolled patients, people with CYP3A5 A6986G GG genotype showed a statistically higher DCR than those with AG + GG genotype (77.8% versus 57.4%, P < 0.05), and the DCR of the patients with GSTP1 A313G AG + GG was statistically higher than that of patients with AA (81.6% versus 57.4%, P < 0.05). Combination analysis showed that patients with GSTP1 A313G AG + GG and CYP3A5 A6986G GG had the highest DCR (84.2%, P < 0.05). CONCLUSION: CYP3A5 and GSTP1 polymorphisms are associated with the short-term efficacy of docetaxel plus thiotepa. Higher DCRs were seen in patients carrying GSTP1 A313G AG + GG and / or CYP3A5 A6986G GG genotype, which might be a reference for clinical chemotherapy. Copyright 2012 by the Chinese Pharmaceutical Association.

Safety Evaluation of Fucoidan from <i>Kjellmaniella Crassiforia</i> and Extract from <i>Hypsizigus Marmoreus</i>: Influence on the Activities and Gene Expressions of Rat Hepatic CYPs / 日本補完代替医療学会誌

<b>Object:</b> Gagome kombu (<i>Kjellmaniella cracciforia</i>) is the edible brown seaweed and contains fucoidan, a sulfated polysaccharide, abundantly. Bunashimeji (<i>Hypsizigus marmoreus</i>) is the popular Japanese mushrooms and contains polyterpenes as the bitter substance. Previously, we investigated the bioactive functions (e.g. anti-tumor action) and the safety of fucoidan from Gagome kombu (GKF) and the extract from Bunashimeji (KTE: Kinoko terpene extract). In this study, we evaluate the influence of GKF and KTE on hepatic cytochrome P450 (CYP).<br> <b>Methods:</b> Male SD rats were divided into three groups (n = 5). 2,000 mg/kg of GKF and KTE were given orally once daily for 4 days.<br> <b>Result:</b> There were no difference in activities and mRNA expressions of hepatic CYPs (CYP2C11, CYP2D, CYP2E1 and CYP3A1) among all groups.<br> <b>Conclusion:</b> These results indicated GKF and KTE did not influence the rat hepatic CYPs.<br>

Safety Evaluation of Extract from Cultured Lentinula edodes Mycelia; Study of Acute Toxicity, Genotoxicity and Inhibiting Effect of Drug-Metabolizing Enzyme, Cytochrome P-450 3A4 / 日本補完代替医療学会誌

<i>Objective</i>: Extract from cultured <i>Lentinula edodes</i> mycelia (L.E.M.) is a food ingredient possessing various pharmacologic actions such as immunomodulatory properties, antitumor and hepatoprotective effects. In Japan, it has been used as a health food for 30 years or more.<br> In the present study to evaluate the safety of L.E.M., a genotoxicity study and acute toxicity study were conducted. In addition, the inhibitory effect of drug-metabolizing enzyme by L.E.M. was tested<i> in vitro</i>, to gain insight on the interaction with medicines.<br> <i>Methods</i>: The genotoxicity study was performed using a bacterial reverse mutation assay and a <i>in vivo</i> mammalian bone marrow cell chromosomal mutation assay. The acute toxicity study was performed using a single-dose oral toxicity test in rats. Inhibitory activity of cytochrome P-450 3A4 (CYP3A4), one of the most important drug-metabolizing enzymes, by L.E.M. was tested using a baculovirus-expressed system.<br> <i>Results</i>: In the genotoxicity study, mutagenicity was negative for both bacterial reverse mutation assay and <i>in vivo</i> mammalian bone marrow cell chromosomal mutation assay. In the acute toxicity study, no toxic symptoms were observed by single dose oral administration of L.E.M. at a dose of 10,000 mg/kg BW in rats. This implies LD₅₀>10,000 mg/kg BW. No inhibitory activity of CYP3A4 by L.E.M. was observed at in the <i>in vitro</i> screening system to investigate drug-L.E.M. interaction.<br> <i>Conclusion</i>: It is believed L.E.M. is a safety ingredient for foods used in complementary and alternative medicine, since it was toxicologically safe and showed no inhibitory activity of CYP3A4 in the studies conducted.<br>

The Effect of All-Trans-Retinoic Acid on the Activity and the Gene Expression of Drug Metabolizing Enzymes in Rat Skin / 대한피부과학회지

BACKGROUND: The activity of drug metabolizing enzymes and the modulation of their expression by inducers in the skin are the key factors for understanding of pharmacological and toxic effects of topically applied drugs. The role of these enzymes is of major importance, as they may contribute to determine the steady-state levels of biologically active substances. 3-Methylcholanthrene and all-trans- retinoic acid have been known to be inducers of the drug metabolizing enzymes. And all-trans- retinoic acid has many biological actions including anti-cancer effects. OBJECTIVE: The purpose of this study was to evaluate the effect of all-trans-retinoic acid on inducing the expression and modulation of genetic polymorphism of drug metabolizing enzymes as well as to estimate the role of all-trans-retinoic acid in carcinogenesis and drug interactions. METHODS: We analyzed the activities of CYP1A1(Cytochrome P450 1Al), NADPH cytochrome P450-reductase, UGT1 and GST after administration of 3-methylcholanthrene and all-trans-retinoic acid to the Sprague-Dawley male rats. We observed the inducible gene expression of CYP1A1, UGT1, GSTJt by RT-PCR and the genetic polymorphism of CYP1A1, UGT1, GSTK by PCR. RESULTS: 1. The expression of CYP1A1, NADPH cytochrome P450-reductase, UGT1 and GST was induced by 3-methylcholanthrene and all-trans-retinoic acid. That of NADPH cytochrome P450-reductase and UGT1 is pronouncedly enhanced by all-trans- retinoic acid. 2. The effects of 3-methylcholanthrene and all-trans-retinoic acid on inducing the expression of CYP1A1 and UGT1 correlated with an increase of mRNA expression levels of CYP1A1 and UGT1. The modulation of mRNA expression levels of GST was downregulated by all-trans-retinoic acid. 3. The genetic polymorphism of CYP1A1 was induced by 3-methylcholanthrene and all-trans- retinoic acid, and that of GSTM1 was not affected by the inducers. The induction of genetic polymorphism of GST was down regulated by all-trans-retinoic acid. CONCLUSION: 3-Methylcholanthrene and all-trans-retinoic acid modulate the inducible expression and genetic polymorphism of drug metabolizing enzymes differentially. All-trans-retinoic acid can modulate the metabolism of procarcinogen such as 3-methylcholanthrene by inducing drug metabolizing enzymes. Furthermore, the elucidation of the molecular mechanisms underlying the regulation of drug metabolizing enzymes by 3-methylcholanthrene, all-trans-retinoic acid and other drugs could help to understand their respective roles in drug interactions and carcinogenesis.

Cytochrome P-450, Drug Metabolizing Enzymes and Arachidonic Acid Metabolism in Pig Ocular Tissues

The eye is a relatively isolated organ and tends to retain or accumulate compounds within itself. Unless the eye possesses a mechanism for detoxifying drugs and chemicals that are carried to ocular tissues via the circulating blood, the compounds may cause visual impairment. The purposes of this experiment was judged by drug metabolizing action by cytochrome P-450 system, distribution and effect of metabolism of arachidonic acid and distribution of lipid peroxide formation and phospholipids in the lens, cornea and choroid-retina of pig ocular tissues. The results were as follows: 1. The activities of mixed-function oxidase, cytochrome P-450 and NADPH-cytochrome C reductase were the highest in the choroid-retina compared to the lens and cornea. 2. The activities of drug metabolizing enzymes, aryl hydrocarbon hydroxylase, 7-ethoxycoumarin-o-deethy lase and benzphetamine-N-demethy lase, heme oxygenase activity, and the activity of arachidonic acid metabolizing enzyme, cyclooxygenase were the highest in the choroid-retina compared to the lens and cornea. 3. Lipid peroxide formation was the highest in the cornea compared to the lens and choroid retina. 4. In distribution of phospholipids, sphingomyelin and phosphatidylcholine were the highest in the choroid-retina compared to the lens and cornea and phosphatidylethanolamine was the highest in the cornea campa red to the lens and choroid-retina. 5. Cytochrome P-450 isoenzyme of pig ocular tissues was confirmed in the same form of MAb(Monoclonal Antibody) PCN(pregenolone-16alpha-carbonitrile) 213-1.